Journal ArticleDOI
Neural differentiation of mouse embryonic stem cells as a tool to assess developmental neurotoxicity in vitro.
Anke Visan,Katrin Hayess,Dana Sittner,Elena E. Pohl,Elena E. Pohl,Christian Riebeling,Birgitta Slawik,Konrad Gulich,Michael Oelgeschläger,Andreas Luch,Andrea Seiler +10 more
Reads0
Chats0
TLDR
The current study shows that D3 mESCs differentiate efficiently into neural cells involving a neurosphere-like state and that this system is suitable to detect adverse effects of neurodevelopmental toxicants.Abstract:
Mouse embryonic stem cells (mESCs) represent an attractive cellular system for in vitro studies in developmental biology as well as toxicology because of their potential to differentiate into all fetal cell lineages. The present study aims to establish an in vitro system for developmental neurotoxicity testing employing mESCs. We developed a robust and reproducible protocol for fast and efficient differentiation of the mESC line D3 into neural cells, optimized with regard to chemical testing. Morphological examination and immunocytochemical staining confirmed the presence of different neural cell types, including neural progenitors, neurons, astrocytes, oligodendrocytes, and radial glial cells. Neurons derived from D3 cells expressed the synaptic proteins PSD95 and synaptophysin, and the neurotransmitters serotonin and γ-aminobutyric acid. Calcium ion imaging revealed the presence of functionally active glutamate and dopamine receptors. In addition, flow cytometry analysis of the neuron-specific marker protein MAP2 on day 12 after induction of differentiation demonstrated a concentration dependent effect of the neurodevelopmental toxicants methylmercury chloride, chlorpyrifos, and lead acetate on neuronal differentiation. The current study shows that D3 mESCs differentiate efficiently into neural cells involving a neurosphere-like state and that this system is suitable to detect adverse effects of neurodevelopmental toxicants. Therefore, we propose that the protocol for differentiation of mESCs into neural cells described here could constitute one component of an in vitro testing strategy for developmental neurotoxicity.read more
Citations
More filters
Journal ArticleDOI
In vitro models of medulloblastoma: Choosing the right tool for the job
TL;DR: Novel ways of improving the clinical relevance of in vitro models are reviewed, focusing on 3D cell culture, extracellular matrix, co-cultures with normal cells and organotypic slices.
Journal ArticleDOI
Recommendation on test readiness criteria for new approach methods in toxicology: Exemplified for developmental neurotoxicity
Anna Bal-Price,Helena T. Hogberg,Kevin M. Crofton,Mardas Daneshian,Rex E. FitzGerald,Ellen Fritsche,Tuula Heinonen,Susanne Hougaard Bennekou,Stefanie Klima,Aldert H. Piersma,Magdalini Sachana,Timothy J. Shafer,Andrea Terron,Florianne Monnet-Tschudi,Barbara Viviani,Tanja Waldmann,Remco H.S. Westerink,Martin F. Wilks,Hilda Witters,Marie-Gabrielle Zurich,Marcel Leist +20 more
TL;DR: Suggestions are made on how DNT NAMs may be assembled into an integrated approach to testing and assessment (IATA), and a vision is presented on how further NAM development may be guided by knowledge of signaling pathways necessary for brain development, DNT pathophysiology, and relevant adverse outcome pathways (AOP).
Journal ArticleDOI
In vitro acute and developmental neurotoxicity screening : an overview of cellular platforms and high-throughput technical possibilities
Béla Z. Schmidt,Martin Lehmann,Simon Gutbier,Erastus Nembu Nembo,Sabrina Noël,Lena Smirnova,Anna Forsby,Juergen Hescheler,Hasan X. Avci,Thomas Hartung,Marcel Leist,Julianna Kobolák,Andras Dinnyes +12 more
TL;DR: The main cellular characteristics underlying neurotoxicity are summarized, an overview of cellular platforms and read-out combinations assessing distinct parts of acute and developmental neurotoxicology are presented, and the use of stem cell-based test systems to close gaps in the available battery of tests is highlighted.
Journal ArticleDOI
Lead exposure disrupts global DNA methylation in human embryonic stem cells and alters their neuronal differentiation.
Marie Claude Senut,Arko Sen,Pablo Cingolani,Pablo Cingolani,Asra N. Shaik,Susan Land,Douglas M. Ruden +6 more
TL;DR: It is shown that exposure to Pb subtly alters the neuronal differentiation of exposed hESCs and that these changes could be partly mediated by modifications in the DNA methylation status of genes crucial to brain development.
Journal ArticleDOI
Human Ntera2 cells as a predictive in vitro test system for developmental neurotoxicity
TL;DR: It is concluded that NT2 cell tests are likely to become valuable components of a human cell-based modular in vitro DNT test systems to assess DNT potential of chemicals at an early stage of brain development.
References
More filters
Journal ArticleDOI
Genome-wide atlas of gene expression in the adult mouse brain
Ed S. Lein,Michael Hawrylycz,Nancy Ao,Mikael Ayres,Amy Bensinger,Amy Bernard,Andrew F. Boe,Mark S. Boguski,Mark S. Boguski,Kevin S. Brockway,Emi J. Byrnes,Lin Chen,Li Chen,Tsuey-Ming Chen,Mei Chi Chin,Jimmy Chong,Brian E. Crook,Aneta Czaplinska,Chinh Dang,Suvro Datta,Nick Dee,Aimee L. Desaki,Tsega Desta,Ellen Diep,Tim A. Dolbeare,Matthew J. Donelan,Hong-Wei Dong,Jennifer G. Dougherty,Ben J. Duncan,Amanda Ebbert,Gregor Eichele,Lili K. Estin,Casey Faber,Benjamin A.C. Facer,Rick Fields,Shanna R. Fischer,Tim P. Fliss,Cliff Frensley,Sabrina N. Gates,Katie J. Glattfelder,Kevin R. Halverson,Matthew R. Hart,John G. Hohmann,Maureen P. Howell,Darren P. Jeung,Rebecca A. Johnson,Patrick T. Karr,Reena Kawal,Jolene Kidney,Rachel H. Knapik,Chihchau L. Kuan,James H. Lake,Annabel R. Laramee,Kirk D. Larsen,Christopher Lau,Tracy Lemon,Agnes J. Liang,Ying Liu,Lon T. Luong,Jesse Michaels,Judith J. Morgan,Rebecca J. Morgan,Marty Mortrud,Nerick Mosqueda,Lydia Ng,Randy Ng,Geralyn J. Orta,Caroline C. Overly,Tu H. Pak,Sheana Parry,Sayan Dev Pathak,Owen C. Pearson,Ralph B. Puchalski,Zackery L. Riley,Hannah R. Rockett,Stephen A. Rowland,Joshua J. Royall,Marcos J. Ruiz,Nadia R. Sarno,Katherine Schaffnit,Nadiya V. Shapovalova,Taz Sivisay,Clifford R. Slaughterbeck,Simon Smith,Kimberly A. Smith,Bryan I. Smith,Andy J. Sodt,Nick N. Stewart,Kenda-Ruth Stumpf,Susan M. Sunkin,Madhavi Sutram,Angelene Tam,Carey D. Teemer,Christina Thaller,Carol L. Thompson,Lee R. Varnam,Axel Visel,Axel Visel,Ray M. Whitlock,Paul Wohnoutka,Crissa K. Wolkey,Victoria Y. Wong,Matthew J.A. Wood,Murat B. Yaylaoglu,Rob Young,Brian L. Youngstrom,Xu Feng Yuan,Bin Zhang,Theresa A. Zwingman,Allan R. Jones +109 more
TL;DR: An anatomically comprehensive digital atlas containing the expression patterns of ∼20,000 genes in the adult mouse brain is described, providing an open, primary data resource for a wide variety of further studies concerning brain organization and function.
Journal ArticleDOI
The in vitro development of blastocyst-derived embryonic stem cell lines: formation of visceral yolk sac, blood islands and myocardium
TL;DR: The in vitro developmental potential of mouse blastocyst-derived embryonic stem cell lines and the consistency with which the cells express this potential are presented as aspects which open up new approaches to the investigation of embryogenesis.
Journal ArticleDOI
Directed Differentiation of Embryonic Stem Cells into Motor Neurons
TL;DR: It is shown that developmentally relevant signaling factors can induce mouse embryonic stem cells to differentiate into spinal progenitor cells, and subsequently into motor neurons, through a pathway recapitulating that used in vivo.
Journal ArticleDOI
Dopamine neurons derived from embryonic stem cells function in an animal model of Parkinson's disease
Jong-Hoon Kim,Jonathan M. Auerbach,José A. Rodríguez-Gómez,Iván Velasco,Denise K. Gavin,Nadya Lumelsky,Sang-Hun Lee,John Bang Nguyen,Rosario Sanchez-Pernaute,Krys S. Bankiewicz,Ronald D.G. McKay +10 more
TL;DR: It is shown that a highly enriched population of midbrain neural stem cells can be derived from mouse ES cells and the dopamine neurons generated by these stem cells show electrophysiological and behavioural properties expected of neurons from the midbrain.
Journal ArticleDOI
Conversion of embryonic stem cells into neuroectodermal precursors in adherent monoculture.
TL;DR: It is reported that neither multicellular aggregation nor coculture is necessary for ES cells to commit efficiently to a neural fate and this system provides a platform for defining the molecular machinery of neural commitment and optimizing the efficiency of neuronal and glial cell production from pluripotent mammalian stem cells.
Related Papers (5)
Developmental neurotoxicity of industrial chemicals
Human embryonic stem cell-derived test systems for developmental neurotoxicity: a transcriptomics approach
Anne K. Krug,Raivo Kolde,John Antonydas Gaspar,Eugen Rempel,Nina V. Balmer,Kesavan Meganathan,Kinga Vojnits,Mathurin Baquié,Tanja Waldmann,Roberto Ensenat-Waser,Smita Jagtap,Richard Evans,Stéphanie Julien,Hedi Peterson,Dimitra Zagoura,Suzanne Kadereit,Daniel Gerhard,Isaia Sotiriadou,Michael Heke,Karthick Natarajan,Margit Henry,Johannes Winkler,Rosemarie Marchan,Luc Stoppini,Sieto Bosgra,Joost Westerhout,Miriam Verwei,Jaak Vilo,Andreas Kortenkamp,Jürgen Hescheler,Ludwig A. Hothorn,Susanne Bremer,Christoph van Thriel,Karl-Heinz Krause,Jan G. Hengstler,Jörg Rahnenführer,Marcel Leist,Agapios Sachinidis +37 more
Critical periods of vulnerability for the developing nervous system: evidence from humans and animal models.
Deborah C. Rice,Stan Barone +1 more